ADAR-mediated RNA editing suppresses sleep by acting as a brake on glutamatergic synaptic plasticity.

It has been postulated that synaptic potentiation during waking is offset by a homoeostatic reduction in net synaptic strength during sleep. However, molecular mechanisms to support such a process are lacking. Here we demonstrate that deficiencies in the RNA-editing gene Adar increase sleep due to synaptic dysfunction in glutamatergic neurons in Drosophila. Specifically, the vesicular glutamate transporter is upregulated, leading to over-activation of NMDA receptors, and the reserve pool of glutamatergic synaptic vesicles is selectively expanded in Adar mutants. Collectively these changes lead to sustained neurotransmitter release under conditions that would otherwise result in synaptic depression. We propose that a shift in the balance from synaptic depression towards synaptic potentiation in sleep-promoting neurons underlies the increased sleep pressure of Adar-deficient animals. Our findings provide a plausible molecular mechanism linking sleep and synaptic plasticity.

A novel electroencephalography-based tool for objective assessment of network dynamics activated by nociceptive stimuli.

BACKGROUND: Pain perception is typically assessed using subjective measures; an objective measure of the response to pain would be valuable. In this study, Brain Network Activation (BNA), a novel multivariate pattern analysis and scoring algorithm, was applied to event-related potentials (ERPs) elicited by cortical responses to brief heat stimuli. Objectives of this study were to evaluate the utility of BNA as a quantitative and qualitative measure of cortical response to pain. METHODS: Contact Heat Evoked Potentials (CHEPs) data were collected from 17 healthy, right-handed volunteers (10 M, 7F) using 5 different temperatures (35, 41, 46, 49 and 52 °C). A set of spatio-temporal activity patterns common to all the subjects in the group (Reference Brain Network Model; RBNM) was generated using the BNA algorithm, based on evoked responses at 52 °C. RESULTS: Frame by frame 'unfolding' of the brain network across time showed qualitative differences between responses to painful and non-painful stimuli. Brain network activation scores were shown to be a better indicator of the individual's sensitivity to pain when compared to subjective pain ratings. Additionally, BNA scores correlated significantly with temperature, demonstrated good test-retest reliability, as well as a high degree of sensitivity, specificity and accuracy in correctly categorizing subjects who reported stimuli as painful. CONCLUSIONS: These results may provide evidence that the multivariate analysis performed with BNA may be useful as a quantitative, temporally sensitive tool for assessment of pain perception.

Mutations in the beta-propeller domain of the Drosophila brain tumor (brat) protein induce neoplasm in the larval brain.

Inactivation of both alleles of the fruit fly D. melanogaster brain tumor (brat) gene results in the production of a tumor-like neoplasm in the larval brain, and lethality in the larval third instar and pupal stages. We cloned the brat gene from a transposon-tagged allele and identified its gene product. brat encodes for an 1037 amino acid protein with an N-terminal B-boxl zinc finger followed by a B-box2 zinc finger, a coiled-coil domain, and a C-terminal beta-propeller domain with six blades. All these motifs are known to mediate protein-protein interactions. Sequence analysis of four brat alleles revealed that all of them are mutated at the beta-propeller domain. The clustering of mutations in this domain strongly suggests that it has a crucial role in the normal function of Brat, and defines a novel protein motif involved in tumor suppression activity. The brat gene is expressed in the embryonic central and peripheral nervous systems including the embryonic brain. In third instar larva brat expression was detected in the larval central nervous system including the brain and the ventral ganglion, in two glands - the ring gland and the salivary gland, and in parts of the foregut - the gastric caecae and the proventriculus. A second brat-like gene was found in D. melanogaster, and homologs were identified in the nematode, mouse, rat, and human. Accumulated data suggests that Brat may regulate proliferation and differentiation by secretion/transport-mediated processes.

Antifungal rapamycin analogues with reduced immunosuppressive activity.

Several 1,2,3,4-tetrahydro- and 7-N-hydroxycarbamate derivatives of the natural product rapamycin were prepared and assayed for their immunosuppressive and antifungal profiles. Substitutions at the 7-position indicate the possibility of a differentiated immunosuppressive to antifungal profile, whereas 40-position variants of the tetrahydro-analogues did not show similar differentiated activity.

Elevated tumor necrosis factor-alpha activation of human immunodeficiency virus type 1 subtype C in Southern Africa is associated with an NF-kappaB enhancer gain-of-function.

The human immunodeficiency virus type 1 (HIV-1) epidemic within southern Africa is predominantly associated with the HIV-1C subtype. Functional analysis of the enhancer region within the long terminal repeat (LTR) indicates that HIV-1C isolates have >/=3 NF-kappaB binding sites, unlike other subtypes, which have only 1 or 2 sites. A correlation was shown between NF-kappaB enhancer configuration and responsiveness to the proinflammatory cytokine tumor necrosis factor (TNF)-alpha within the context of naturally occurring subtype LTRs, subtype-specific NF-kappaB enhancer regions cloned upstream of an isogenic HXB2 core promoter or a heterologous SV40 minimal promoter, and full-genome subtype clones. In all cases, TNF-alpha activation was correlated with the subtype configuration of the NF-kappaB enhancer. Whether the naturally occurring gain-of-function in the NF-kappaB enhancer of HIV-1C observed in this study can provide a selective advantage for the virus in vivo remains to be determined and warrants further study.

Potent and orally bioavailable noncysteine-containing inhibitors of protein farnesyltransferase.

Potent and orally bioavailable nonthiol-containing inhibitors of protein farnesyltransferase are described. Oral bioavailability was achieved by replacement of the pyridyl ether moiety of 1 with a 2-substituted furan ether to give 4. Potency was regained with 2,5-disubstituted furan ethers while maintaining the bioavailability inherent in 4. p-Chlorophenylfuran ether 24 is 0.7 nM in vitro (FTase) and is 32% bioavailable in the mouse, 30% bioavailable in rats, and 21% bioavailable in dogs.

Organization resources for families of children with deafness or hearing loss.

National health and education mandates emphasize the value of and need for early identification and intervention for children who are deaf and hard of hearing. Pediatricians in collaboration with other professionals in a multidisciplinary approach play a vital role in linking families to appropriate care. Understanding the concept of choices in communication and education resources and the process of parental and caregiver grieving enables the pediatrician to assist the family in finding needed services, agencies, and resources.

Protein synthesis elongation factor EF-1 alpha is an isopeptidase essential for ubiquitin-dependent degradation of certain proteolytic substrates.

Targeting of different cellular proteins for conjugation and subsequent degradation via the ubiquitin pathway involves diverse recognition signals and distinct enzymatic factors. A few proteins are recognized via their N-terminal amino acid residue and conjugated by a ubiquitin-protein ligase that recognizes this residue. However, most substrates, including N-alpha-acetylated proteins that constitute the vast majority of cellular proteins, are targeted by different signals and are recognized by yet unknown ligases. In addition to the ligases, other factors may also be specific for the recognition of this subset of proteins. We have previously shown that degradation of N-terminally blocked proteins require a specific factor, designated FH, and that the factor acts along with the 26S protease complex to degrade ubiquitin-conjugated proteins (Gonen et al., 1991). Further studies have shown that FH is identical to the protein synthesis elongation factor EF-1 alpha, and that it can be substituted by the bacterial elongation factor EF-Tu (Gonen et al., 1994). This, rather surprising, finding raises two important and interesting problems. The first involves the mechanism of action of the factor and the second the possibility that protein synthesis and degradation may be regulated by a commonly shared factor. Here, we demonstrate that EF-1 alpha is a ubiquitin C-terminal hydrolase (isopeptidase) that is probably involved in trimming the conjugates to lower molecular weight forms recognized by the 26S proteasome complex. Additional findings demonstrate that its activity is inhibited specifically by tRNA. This finding raises the possibility that under anabolic conditions, when the factor is associated with AA.tRNA and GTP, it is active in protein synthesis but inactive in proteolysis. Under catabolic conditions, when the factor is predominantly found in its apo form, it is active in proteolysis.

Class II cytoplasmic and transmembrane domains are not required for class II-mediated B cell spreading.

B cells cultured on immobilized anti-class II monoclonal antibody (mAb) change from round to flattened cells, with lamellipodia and filopodia. This change in cell morphology, termed 'spiders', occurs within 30 min upon culture and is mediated through either I-A or I-E molecules. Class II molecules that are defective in mediating protein kinase C (PKC) due to the deletions of both alpha and beta chain's cytoplasmic (Cy) domain sequences can induce spider formation. B-cell transfectants that express chimeric MHC class II/class I molecules, where the ectodomains are class II sequences and the transmembrane and Cy domains are class I sequences also form spiders when cultured on anti-class II mAb. The spider morphology is not induced by either anti-immunoglobulin (Ig) or anti-MHC class I mAb. Treatment of B cells to increase intracellular cAMP, a component of the class II signaling pathway also results in spider formation with the same kinetics and percent change in the responding population as that induced by anti-class II mAb. Cytochalasin A treatment which disrupts cytoskeletal actin filaments and the tyrosine kinase inhibitor, genistein, both inhibit spider formation. Actin redistributes from a concentric ring in round cells to the ends of the filopodia in the spiders. The mechanism of spider induction whether resultant from second messengers following class II signaling or from non-signaling-induced physical interactions of class II with intracellular cytoskeletal components only requires the extracellular domains of class II. The biologic relevance of B-cell spiders is currently not known but has been reported to be associated with class II signal transduction and efficient Ag presentation.

Gestational age assessment.

Accurate assessment of gestational age is an essential component of good obstetric care. Many techniques of gestational estimation are available, ranging from calculations based on the date of the last normal menstrual period to sonographic measurements of fetal parameters. Each method has advantages and disadvantages and a stage of pregnancy at which it is most reliable. Early attention to appropriate indicators, reinforced by the monitoring of gestational parameters throughout pregnancy, leads to a more accurate estimated date of confinement than reliance on a single indicator. Incongruity between fetal size and dates should arouse suspicion of specific, serious clinical problems.

Use of nutrient response techniques to assess the effectiveness of chlorination of rapid sand filter gravel.

A direct viable counting method was used to rapidly assess the effectiveness of chlorination of biofilms on rapid sand filter gravel. A total of 50% of the cells were nutrient responsive after exposure to 0.5 mg of chlorine per liter, while this value was 25% after exposure to 25 mg of chlorine per liter. A large variation was seen in the numbers of nutrient-responsive cells on different rocks. More cells attached to the sandblasted side of marbles than to the smooth side, but there was no difference in eight of nine cases in the proportion of survival to chlorination between the two different sides. The effectiveness of chlorination appeared to be influenced by the species of bacterium in the biofilm.

Comparison of methods for enumeration of selected coliforms exposed to ozone.

mT7 medium performed no better than m-Endo medium in enumerating cells of Escherichia coli and Citrobacter freundii exposed to ozone. Also, there was no difference in the plate count of heterotrophic bacteria in ozonated raw water determined on modified Henrici agar or R2A agar. Statistically significant differences were seen between bacteria and the type of water in which they were suspended during ozonation.

Integrating systems theory and attachment theory: the use of radio communication to modify attachments in a patient with psychogenic vomiting.

A case is presented that shows the usefulness of integrating systems theory and attachment theory in the formulation and treatment of a clinical problem. The 5 year old son of East Indian immigrants presented with persistent psychogenic vomiting associated with pathological family attachments. It was evident that the precarious family equilibrium was stabilized by the child's psychogenic vomiting. The therapeutic team suggested to the family that their problems might be more satisfactorily resolved if the mother and child maintained their link by two-way radio. Three weeks later the vomiting had ceased, the child no longer felt that he needed the radio and both parents had established new patterns of relating to their child, whose attendance and peer socialization at school showed marked improvement. To some extent the rapid resolution of the problems was facilitated by the cultural strengths of the family.

Sensitivity to thermal stimulation in prairie rattlesnakes (Crotalus viridis) after bilateral anesthetization of the facial pits.

Six yearling prairie rattlesnakes (Crotalus viridis) were exposed to thermal stimuli prior to and after bilateral anesthetization of their facial pits with 2% xylocaine solution. This treatment eliminates trigeminally mediated electrophysiological responses of the pits to thermal stimulation. Nevertheless, the rattlesnakes continued to exhibit behavioral responses to thermal cues after anesthetization of the pits. An auxiliary infrared-sensitive system, nociceptors, or the common temperature sense could be responsible for these findings.

Susceptibility of clinical isolates of Campylobacter jejuni to sixteen antimicrobial agents.

The activities of 16 antimicrobial agents against 103 clinical isolates of Campylobacter jejuni were tested. All the strains were susceptible to kanamycin and gentamicin. Chloramphenicol, nalidixic acid, and clindamycin were active against most of the strains. More than one-third of the strains were resistant to the tetracyclines and 12.5% were resistant to erythromycin.

Serotype distribution of penicillin-resistant pneumococci and their susceptibilities to seven antimicrobial agents.

A total of 229 clinical isolates of Streptococcus pneumoniae recovered from 225 patients were serotyped and tested for susceptibility to penicillin G, ampicillin, mezlocillin, cefazolin, erythromycin, clindamycin, chloramphenicol, and sulfamethoxazole-trimethoprim. Of all the isolates, 48 (21.0%) showed intermediate resistance and 17 (7.4%) showed resistance to penicillin G. Penicillin-resistant strains had higher minimal inhibitory concentrations of ampicillin, mezlocillin, and cefazolin than did penicillin-susceptible strains. Resistance to erythromycin and clindamycin was rare (1.3 and 0.9%, respectively). Of the isolates, 8.7% were resistant to sulfamethoxazole-trimethoprim, and all were susceptible to chloramphenicol. Penicillin resistance was associated with 13 serotypes. Serotypes 14, 19F, 19A, and 23F were both highly prevalent and frequently penicillin resistant.

A simple EMG--reward system for biofeedback training of children.

An automated reward system to be used with standard electromyographic (EMG) biofeedback instrumentation is described. The on-line reward system, when coupled with standard biofeedback instrumentation and procedures, constitutes instrumental conditioning. In addition to describing the EMG--reward interface circuitry, a study demonstrating the efficacy of the instrumental conditioning approach with cerebral-palsied children is presented.